Cohen, E. A. K. et al. Nat. Commun. 10, 793 (2019).
Single-molecule localization microscopy methods such as photoactivated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM) rely on image-processing algorithms that identify the positions of individual fluorescent emitters in a given frame en route to full image reconstruction. The resolution of these images is intricately linked to algorithm performance, yet no approaches have been available to evaluate the resolving capabilities of such algorithms. Cohen et al. addressed this challenge by using spatial statistics to explore the impact of algorithm performance on resolution. They found that achievable resolution varies widely among the tools tested, with some being much larger than diffraction-limited resolution. They also show that these differences can affect downstream analysis, such as the quantitative analysis of protein clustering. However, they show that incorporation of the known algorithmic resolution can be used to correct clustering analysis for accurate results.
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Strack, R. On algorithmic resolution. Nat Methods 16, 285 (2019). https://doi.org/10.1038/s41592-019-0383-1